한빛사논문
Kibaek Choe1,*, Yusaku Hontani 1,6, Tianyu Wang 1, Eric Hebert1, Dimitre G. Ouzounov1, Kristine Lai2, Ankur Singh2,3,4, Wendy Béguelin 5, Ari M. Melnick5 and Chris Xu 1,*
1School of Applied and Engineering Physics, Cornell University, Ithaca, NY, USA. 2School of Mechanical Engineering, College of Engineering, Cornell University, Ithaca, NY, USA. 3Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, USA. 4Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA, USA. 5Division of Hematology/Oncology, Department of Medicine, Weill Cornell Medical College, Cornell University, New York, NY, USA. 6Present address: Brain Research Institute, University of Zurich, Zurich, Switzerland.
*Correspondence to Kibaek Choe or Chris Xu.
Abstract
Intravital confocal microscopy and two-photon microscopy are powerful tools to explore the dynamic behavior of immune cells in mouse lymph nodes (LNs), with penetration depth of ~100 and ~300 μm, respectively. Here, we used intravital three-photon microscopy to visualize the popliteal LN through its entire depth (600–900 μm). We determined the laser average power and pulse energy that caused measurable perturbation in lymphocyte migration. Long-wavelength three-photon imaging within permissible parameters was able to image the entire LN vasculature in vivo and measure CD8+ T cells and CD4+ T cell motility in the T cell zone over the entire depth of the LN. We observed that the motility of naive CD4+ T cells in the T cell zone during lipopolysaccharide-induced inflammation was dependent on depth. As such, intravital three-photon microscopy had the potential to examine immune cell behavior in the deeper regions of the LN in vivo.
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